Results:
a. b. c.
Figure 1) a. Plate showing colony growth on Agar plate with ampicillin. The bacteria grown on this plate is BL21(DE3). The plasmid within the bacteria is pEM-gbr22. The colonies are small whitish-yellow dots on the Agar on the plate. b. Plate showing no colony growth on Agar with ampicillin. The bacteria on this plate is BL21(DE3). There is no plasmid in the bacteria on this plate. There are no colonies shown on this plate. c. Plate showing no colony growth on Agar with no ampicillin. The bateria on this plate is from: Joey's forehead, gloves in the trash bin, men's urinal, men's room paper towel dispenser, mouths (Andee and I - coughs). There is also LB on this plate. No growth is shown on this plate.
Figure 2) Purple culture in flasks. Cultures contain 25 mL of LB Broth, 625 microliters of stater culture, and 50 microliters of ampicillin. The starter culture contains BL21(DE3) bacteria and pGEM-gbr22 plasmid. These pictures were taken after these flasks were shaken in an incubator at 37 degrees Celsius and 200-350 rpm for approximately 24 hours.
Figure3) Cell pellet present in 50 mL conical tube after being centrifuged for 10 minutes at 4 degrees Celsius and 5000 rpm and separated from the liquid that was present. This pellet contains all BL21(DE3) cells containing pGEM-gbr22 plasmid that are expressing the "purple" protein.
Figure 4) 15 mL conical tubes showing approximately 5 mL each of Elution 1 on the bottom and Elution 2 on the top, respectively. The elutions contain the purified protein and elution buffer.
Figure 5) NanoDrop results for a sample of Elution 1 set at 280 nm under the A280 setting. Wavelength in nanometers is shown on the X axis, and 10 mm Absorbance is shown on the Y axis. Concentration reading in mg/ml is shown in the bottom corner.
Figure 6) NanoDrop results for a sample of Elution 1 set at both 280 (on the left) and 574 (on the right) nanometers under the UV-VIS setting. Wavelength in nanometers is shown on the X axis, and the 1 mm Absorbence is shown on the Y axis.
Figure 7) Polyacrylamide Gel showing, from top to bottom, a ladder, bands for the Cell Lysate, bands for the Soluble Fraction, bands for the Flow Through, bands for the Wash, bands for Elution 1, bands for Elution 2, bands for Elution 1 for Daren Nguyen, and bands for Elution 2 for Daren Nguyen. Stained with bromophenol blue. Dried for 1.5 hours.
Introduction:
Materials & Methods:
Results:
a. b. c.
Figure 1) a. Plate showing colony growth on Agar plate with ampicillin. The bacteria grown on this plate is BL21(DE3). The plasmid within the bacteria is pEM-gbr22. The colonies are small whitish-yellow dots on the Agar on the plate. b. Plate showing no colony growth on Agar with ampicillin. The bacteria on this plate is BL21(DE3). There is no plasmid in the bacteria on this plate. There are no colonies shown on this plate. c. Plate showing no colony growth on Agar with no ampicillin. The bateria on this plate is from: Joey's forehead, gloves in the trash bin, men's urinal, men's room paper towel dispenser, mouths (Andee and I - coughs). There is also LB on this plate. No growth is shown on this plate.
Figure 2) Purple culture in flasks. Cultures contain 25 mL of LB Broth, 625 microliters of stater culture, and 50 microliters of ampicillin. The starter culture contains BL21(DE3) bacteria and pGEM-gbr22 plasmid. These pictures were taken after these flasks were shaken in an incubator at 37 degrees Celsius and 200-350 rpm for approximately 24 hours.
Figure3) Cell pellet present in 50 mL conical tube after being centrifuged for 10 minutes at 4 degrees Celsius and 5000 rpm and separated from the liquid that was present. This pellet contains all BL21(DE3) cells containing pGEM-gbr22 plasmid that are expressing the "purple" protein.
Figure 4) 15 mL conical tubes showing approximately 5 mL each of Elution 1 on the bottom and Elution 2 on the top, respectively. The elutions contain the purified protein and elution buffer.
Figure 5) NanoDrop results for a sample of Elution 1 set at 280 nm under the A280 setting. Wavelength in nanometers is shown on the X axis, and 10 mm Absorbance is shown on the Y axis. Concentration reading in mg/ml is shown in the bottom corner.
Figure 6) NanoDrop results for a sample of Elution 1 set at both 280 (on the left) and 574 (on the right) nanometers under the UV-VIS setting. Wavelength in nanometers is shown on the X axis, and the 1 mm Absorbence is shown on the Y axis.
Figure 7) Polyacrylamide Gel showing, from top to bottom, a ladder, bands for the Cell Lysate, bands for the Soluble Fraction, bands for the Flow Through, bands for the Wash, bands for Elution 1, bands for Elution 2, bands for Elution 1 for Daren Nguyen, and bands for Elution 2 for Daren Nguyen. Stained with bromophenol blue. Dried for 1.5 hours.
Discussion:
Conclusions:
References: